New England Biolabs Canada
 

Product Pathways - Chromatin Regulation / Epigenetics

SPT16 (D7I2K) Rabbit mAb #12191

Item# Description List Price Web Price Qty
12191S SPT16 (D7I2K) Rabbit mAb - 100 µl $372.00
$334.80
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Monkey Endogenous 140 Rabbit IgG
ChIP

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, ChIP=Chromatin IP

Homology

Species predicted to react based on 100% sequence homology: Hamster, Xenopus, Zebrafish, Bovine, Dog, Guinea Pig, Horse.

Directions For Use

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

Protocols

Specificity / Sensitivity

SPT16 (D7I2K) Rabbit mAb recognizes endogenous levels of total SPT16 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu662 of human SPT16 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using SPT16 (D7I2K) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT 116 cells starved for 48 hr then serum stimulated with 20% FBS for 15 min and either SPT16 (D7I2K) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human c-Fos Promoter Primers #4663, SimpleChIP® Human c-Fos Exon 3 Primers #12010, SimpleChIP® Human EGR1 Promoter Primers #5549, and SimpleChIP® Human EGR1 Intron 3 Primers #11953. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

Suppressor of Ty-16 (SPT16) and structure-specific recognition protein-1 (SSRP1) are subunits of the facilitates chromatin transcription (FACT) complex that is essential for transcription elongation (1,2). FACT facilitates RNA polymerase-dependent transcription of chromatin templates by destabilizing the nucleosomes within the open reading frames of active genes (3-5). FACT destabilizes the nucleosomes, which would otherwise act as barriers to RNA polymerase transcription activity, by disrupting histone-histone and histone-DNA contacts that lead to the eviction of the histone H2A-H2B dimer (2,3,6). FACT may also function as a histone chaperone to reassemble nucleosomes after RNA polymerase passage (7). In addition to transcription, FACT activity has been shown to have a role in DNA replication in yeast and in DNA repair by contributing to the activation of p53 by CK2 and by facilitating histone H2AX-H2B exchange upon DNA damage (8-10).

  1. Winkler, D.D. and Luger, K. (2011) J Biol Chem 286, 18369-74.
  2. Orphanides, G. et al. (1999) Nature 400, 284-8.
  3. Orphanides, G. et al. (1998) Cell 92, 105-16.
  4. Birch, J.L. et al. (2009) EMBO J 28, 854-65.
  5. Orphanides, G. and Reinberg, D. (2000) Nature 407, 471-5.
  6. Keller, D.M. and Lu, H. (2002) J Biol Chem 277, 50206-13.
  7. Belotserkovskaya, R. et al. (2003) Science 301, 1090-3.
  8. Schlesinger, M.B. and Formosa, T. (2000) Genetics 155, 1593-606.
  9. Keller, D.M. and Lu, H. (2002) J Biol Chem 277, 50206-13.
  10. Heo, K. et al. (2008) Mol Cell 30, 86-97.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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